Fig 5. Induction of phosphorylation of Akt at Thr³⁰⁸ results in IE1 induction during infection with WT HCMV.

(A to C) Monocytes were treated with PBS or GM-CSF (100 ng/ml) for 30 min and mock-infected or infected with WT or US28Δ HCMV (MOI = 1) for 30 min (A) or 24 h (B, C). (D to F) Monocytes were treated with 15 μM of indicated phosphatidylinositol (PIP) or empty lipid carrier (EC) for 50 min and mock-infected or infected with WT and US28Δ HCMV for 30 min (D) or 24 h (E, F). (G to I) Monocytes were transfected with myristyolated-Akt (myr-Akt) plasmid or empty vector (EV) for 48 h and were mock-infected or infected with WT, or US28Δ infected for 30 min (G) or 24 h (H, I). Phosphorylation of Akt at Ser⁴⁷³ and Thr³⁰⁸ and total Akt (A, D, G) and IE1 (B, E, H) were detected by Western blot and quantified (C, F, I). β-actin was used as a loading control. Western blots and densitometry are representative of at least 3 biological replicates per group. *P<0.05, **P < 0.005, ***P < 0.0005, **** P < 0.0001 by one-way ANOVA with Tukey’s HSD post hoc test.